Meta Analysis Of Bovine Viral Diarrhea Virus Epidemic In China And Transcriptome Analysis Of MDBK Cells Infected With BVDV

Bovine viral diarrhoea(BVD)is an infectious disease that causes bovine diarrhea,miscarriage,and stillbirth.It can cause serious losses to the animal husbandry,and the overall prevalence of BVDV in China is still unclear.Meta-analysis can reveal the basic prevalence of BVDV in different regions of China,and estimate potential relevant factors,which will pave the way for identifying the prevalence of the disease in the domestic range of herds.In recent years,most of them are related to BVDV protein research.There are few studies on the level of transcriptome.Because genes infected with BVDV can modify the genes that are favorable for virus survival and regulate related coding genes.Therefore,in order to understand the overall epidemic situation of BVDV in China and the interaction mechanism between host and BVDV,the meta-analysis method was used to evaluate the overall prevalence of BVDV.In addition,high-throughput sequencing methods were used to compare different time periods(0 h,12 h,48 h,and 72 h).The mRNA transcriptome genes were differentially expressed between the challenge cells of 24 and the control group,providing some basic data for elucidating the pathogenic mechanism of the virus.(1)Meta-analysis was used to analyze the BVDV articles published in China from March 15,2003 to March 28,2018.In the Meta analysis,four literature libraries were searched,including China Knowledge Network,Weipu,Wanfang,and PubMed.A total of 1,210 relevant documents were retrieved.Finally,50 articles were selected for quantitative analysis.Based on the data obtained from the 46,801 head of cattle in these documents,the total prevalence rate was 44.59%,and there were different prevalence rates in different regions,which further confirmed the prevalence of BVDV in cattle.According to the criteria of the content of the article,the prevalence of BVDV,the purpose of research,and the improvement of materials and methods,we get 25 papers were of high quality(3 or 4 points),17 papers were of medium quality(2 points),and the remaining 8 papers were of low quality(0 or 1 point);The positive rate in Central China was 66%,which was higher than that in other parts of China.The infection rate in Shaanxi Province was 88.88%.Ningxia was infected.The rate was 97.44%,after 2010 the infection rate was 52% compared to the pre-2010 positive rate,which may be due to the mode of large-scale farming.Development has caused the spread of persistent diseases in livestock farms.(2)A total of 6 G clean data was measured for the transcription spectrum.Among them,43484252 high-quality data were obtained in the control group,45187454 high-quality data were obtained at 12 h,and high-quality data was 46410432 at 24 h,which was high at 48 h.The quality data was 42631906,and 42827480 pieces of high-quality data were obtained at 72 h;the selected differential genes(requiring P value less than 0.05)were compared during the 0 h-12 h infection period,and there were 56 differential genes;after infection,There were 390 differentially expressed genes between 12 h and 24 h,487 differential genes within 24 h to 48 h,and 164 differentially expressed genes between 48 h and 72 h after infection.The information analysis software was used to predict the differentially expressed genes,and the differential genes were analyzed by using GO analysis and KEGG analysis,respectively.GO analysis showed that some genes in BVDV were up-regulated after infecting MDBK cells,most of which were related to cell metabolism and biological regulation,and showed significant enrichment;KEGG analysis showed that the cells infected with virus were in JAK-STAT.Pathways,PI3K/Akt signal transduction pathways,and NF-kappa B pathways showed significant enrichment,and the differentially expressed genes after infection were P21,FOXO1,GABARAPL1,etc.,which were mainly related to cell cycle replication,apoptosis,and immunity.Signal path.(3)Ten differentially expressed genes,including GABARAPL1,CDKN1 A,CCNE2,CDK2,RRM2,ATM,BIRC-5,CDKN1 A,CTSH,and NFKBIA,were selected in the experiment and verified by q-PCR.The results were consistent with the sequencing results.In order to explore the effect of BVDV on apoptosis during different time periods,the changes of PI3 K,AKT and FOXO genes at different time points were verified by q-PCR and western-blot methods from the level of transcription and protein expression.The transcript level increased with the increase of virus infection,and the transcript levels of AKT and FOXO also showed an upward trend.The trend of up-regulation was most obvious at 48 h after infection.The results showed that BVDV activated the PI3 K factor in FOXO signaling pathway after infecting host cells.In turn,activation of the signaling pathway leads to an immune response in the cell;And the results of protein expression levels were consistent with the analysis of mRNA expression levels.As the upregulation of PI3 K can activate the key molecules of the downstream pathway,the results show that the two genes are up-regulated by verifying the levels of downstream genes related to AKT and FOXO,and both show a clear upward trend at 48 h and gradually increase.Through experiments we found a pathway related to apoptosis(FOXO signaling pathway).In summary,the infection rate of BVDV in some parts of China is still very high,and there are some potential related factors that make the disease continue to spread.The results of transcriptome analysis showed that the expression of some differentially expressed genes in MDBK cells was significantly changed in different time periods after BVDV infection.The mRNA and protein levels were verified by PI3 K,AKT and FOXO differential genes,and it was found that the transcription and protein levels increased with the increase of infection time..The exploration of this experiment will pave the way for the subsequent studies of related molecular mechanisms such as BVDV infection.